CPSI (Cell Preservation Services, Inc) is a biotechnology company dedicated to developing novel technologies in the area of cell-molecular biology and medicine. CPSI's core molecular strategy is focused on understanding and manipulating the cell survival and death pathways that appear to be cell-specific and are activated as a consequence of cell manipulation. CPSI's knowledge of these stress pathways has led to insight into the development of a series of future products designed to improve cell bioprocessing. The goal of this research project is to develop the first generation of CellGuardTM, tissue culture additives designed to mitigate cell response to the continually changing environment a cell is exposed to during a bioprocessing procedure. The objective of CellGuardTM is to provide for a more stable culture environment thereby extending the usable interval of cells in vitro while maintaining cell performance within a standard, predictable response window. To this end, CPSI has identified a unique class of compounds, FR-48, that has shown promise. As such, these compounds offer great potential to support CPSI's cell processing program. The intent of this proposal is to (1) better understand the cell stress pathway targeted by FR-48, (2) evaluate FR-48 analogues to assess their efficacy and (3) investigate the utilization of FR-48 analogs in a pre-, intra, and post-manipulation exposure setting. Samples will be evaluated under three distinct environmental stress regimes including hypoxia, nutrient deprivation, and cell growth/aging. Phase 1 studies will utilize the HepG2/C3A liver cell line and primary hepatocytes. Phase 2 studies will be expanded to additional cell types and more in- depth analogue and cell function analysis. Investigations under this Phase 1 will include molecular analysis of FR-48's action using fluorescent assays, flow cytometry, microarray analysis, western blotting, etc. Successful completion of the project will lead to improved protocols for the active, bulk cell culture (bioprocessing) of human hepatocytes as well as other cell systems. Phase 2 studies will focus on the further development and application of the CellGuardTM product line to improve bioprocessing applications for an array of cell systems. PUBLIC HEALTH RELEVANCE: This project is designed to develop an improved series of solutions and protocols for protecting human cells during bioprocessing. This novel approach, collectively called CellGuardTM, will result in the improved viability and function of human cells such as liver cells. This, in turn, will improve the quality of future cell-based, pharmaceutical drugs as well as positively affect the use of human cells in cell therapy.